|本期目录/Table of Contents|

[1]郭兴启,李菡,张杰道,等.表达马铃薯Y病毒外壳蛋白基因的转基因烟草抗病机制[J].应用与环境生物学报,2003,9(04):372-376.
 GUO Xingqi,et al..Evidence for RNA-mediated resistance to PVYn in tobacco plants transformed with the viral coat protein gene[J].Chinese Journal of Applied & Environmental Biology,2003,9(04):372-376.
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表达马铃薯Y病毒外壳蛋白基因的转基因烟草抗病机制()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
9卷
期数:
2003年04期
页码:
372-376
栏目:
综述
出版日期:
2003-08-25

文章信息/Info

Title:
Evidence for RNA-mediated resistance to PVYn in tobacco plants transformed with the viral coat protein gene
作者:
郭兴启李菡张杰道竺晓平王洪刚
Author(s):
GUO Xingqiet al.
( College of Life Sciences; 1 College of Plant Protection, Shandong Agricultural University, Tai an271018, China )
关键词:
关键词 马铃薯Y病毒 外壳蛋白基因 转基因烟草 抗病机制
Keywords:
Keywords Potato virus Y coat protein gene transgenic tobacco resistance mechanism
摘要:
摘要 根据已报道的马铃薯Y病毒坏死株系(PVYN)核苷酸序列,克隆了PVYN外壳蛋白(CP)基因,通过根癌农杆菌(Agrobacterium tumefaciens)LBA4404介导的方法,转化烟草NC89,获得了38株PCR呈阳性的转基因植株.攻毒试验发现,转基因植株对PVYN的抗性水平存在着差异,其中有4株表现为高度抗病性.Southern blot表明,PVYN CP基因已经整合到烟草染色体中,转基因的拷贝数与植株的抗病性成正相关.Northern blot 表明,PVYN CP基因在RNA水平上得到了表达,而且PVYN CP RNA在细胞中的积累量与转基因植株的抗病性呈明显的负相关.Western blot表明,高度抗病植株未检测到转基因CP蛋白质,而抗病和感病植株都检测到了PVYN CP蛋白,且不同抗性的转基因植物中转基因蛋白质的积累有一定的差异.实验结果表明,表达PVYN CP基因的转基因烟草其抗病机制类似于转录后的基因沉默,即抗病性可能是RNA介导的. 图7 表1 参13
Abstract:
Abstract According to the published nucleotide sequence of tobacco veinal necrosis strain (PVYN), the PVYN coat protein (CP) gene was synthesized by reverse transcription-polymerase chain reaction, and constructed into the cloning vector pBSK. The PVYN CP gene was introduced into tobacco (NC89) plants via Agrobacterium tumefaciens (LBA4404)-mediated transformation and the transformed tissue was selected in the presence of Kanamycin sulfate. Regenerated plants were screened by PCR and 38 transgenic plants were obtained. Resistant test indicated that the resistant levels of the transgenic plants against PVYN varied among different transgenic plants. Four lines were highly resistant to PVYN infection. Southern blot analysis indicated that the CP gene had been integrated into the tobacco chromosomes and the resistance levels of the transgenic plants were related to copy numbers of the transgene. The highly resistant plants contained 5~6 copies of the transgene, the resistant plants contained 3~4 copies and the susceptible plants contained 1~2 copies. Northern blot analysis demonstrated that the transgene was expressed at RNA level and there was an inverse correlation between transgene transcript accumulation and virus resistance. Western blot analysis showed that no specific protein was observed in highly resistant plants, transgene CP detected in both the resistant and susceptible plants, and the CP accumulation varied among different transgenic plants. Results indicated that the resistance mechanism of the transgenic plants was similar to post-transcriptional gene silence, i.e., RNA-mediated resistance. Fig 7, Tab 1, Ref 13
更新日期/Last Update: 2001-08-26